Pharmaceutical composition for inhibiting growth of cancers

ABSTRACT

This invention is a pharmaceutical composition that inhibits the growth of cancers and tumors in mammals, particularly in human and warm blooded animals. The composition contains N-chlorophenylcarbamates and N-chlorophenylthiocarbamates along with a chemotherapeutic agent and optionally a potentiator. A composition for treating viral infections in animals or humans comprising a safe and effective amount of N-chlorophenylcarbamates and the N-chlorophenylthiocarbamates and a potentiator is also disclosed.

This is a division of application Ser. No. 08/876,705, filed on Jun. 16,1997 now U.S. Pat. No. 5,932,609 which is a division of Ser. No.08/680,468 filed on Jul. 15, 1996 now U.S. Pat. No. 5,932,604.

TECHNICAL FIELD

This invention is a pharmaceutical composition that inhibits the growthof cancers and tumors in mammals, particularly in human and warm bloodedanimals. The composition is also effective against viruses. Thecomposition contains N-chlorophenylcarbamates andN-chlorophenylthiocarbamates along with potentiators or chemotherapeuticagents or antiviral drugs.

BACKGROUND OF THE INVENTION

Cancers are the leading cause of death in animals and humans. The exactcause of cancer is not known, but links between certain activities suchas smoking or exposure to carcinogens and the incidence of certain typesof cancers and tumors has been shown by a number of researchers.

Many types of chemotherapeutic agents have been shown to be effectiveagainst cancers and tumor cells, but not all types of cancers and tumorsrespond to these agents. Unfortunately, many of these agents alsodestroy normal cells. The exact mechanism for the action of thesechemotherapeutic agents are not always known.

Despite advances in the field of cancer treatment the leading therapiesto date are surgery, radiation and chemotherapy. Chemotherapeuticapproaches are said to fight cancers that are metastasized or ones thatare particularly aggressive. Such cytocidal or cytostatic agents workbest on cancers with large growth factors, i.e., ones whose cells arerapidly dividing. To date, hormones, in particular estrogen,progesterone and testosterone, and some antibiotics produced by avariety of microbes, alkylating agents, and anti-metabolites form thebulk of therapies available to oncologists. Ideally cytotoxic agentsthat have specificity for cancer and tumor cells while not affectingnormal cells would be extremely desirable. Unfortunately, none have beenfound and instead agents which target especially rapidly dividing cells(both tumor and normal) have been used.

Clearly, the development of materials that would target tumor cells dueto some unique specificity for them would be a breakthrough.Alternatively, materials that were cytotoxic to tumor cells whileexerting mild effects on normal cells would be desirable.

Therefore, it is an object of this invention to provide a pharmaceuticalcomposition that is effective in inhibiting the growth of tumors andcancers in mammals with mild or no effects on normal cells.

More specifically, it is an object of this invention to provide ananti-cancer composition comprising a pharmaceutical carrier,chemotherapy agent and a N-chlorophenylcarbamate orN-chlorophenylthiocarbamate derivative as defined herein, along with amethod of treating such cancers.

These compositions are also effective against viruses. Therefore it is afurther object of this invention to provide a composition effectiveagainst HIV, herpes, influenza, rhinoviruses and the like wherein apotentiator is used to improve the effectiveness of the composition.

These and other objects will become evident from the following detaileddescription of this inventions.

SUMMARY OF THE INVENTION

A pharmaceutical composition for treatment of mammals, and inparticular, warm blooded animals and humans, comprising a pharmaceuticalcarrier and an effective amount anti-cancer compound selected from thegroup consisting of N-chlorophenylcarbamates andN-chlorophenylthiocarbamates of the formula:

wherein n is from 1 to 3, X is oxygen or sulfur, and R is selected fromthe group consisting of hydrogen, lower alkyl and lower alkenyl,cyclohexyl, phenalkyl radicals of up to 8 carbon atoms, and phenyl, andpharmaceutically acceptable inorganic or organic acid salts of thesecompounds.

These compositions can be used to inhibit the growth of cancers andother malignant tumors in humans or animals by administration of aneffective amount of the N-chlorophenylcarbamates andN-chlorophenylthiocarbamates either orally, rectally, topically orparenterally, intravenously, or by direct injection near or into thetumor. These compositions are effective in killing or slowing the growthof tumors, yet are safer than adriamycin on normal, healthy cells. Thecompositions are also useful for treating viral infections.

DETAILED DESCRIPTION OF THE INVENTION

A. Definitions:

As used herein, the term “comprising” means various components can beconjointly employed in the pharmaceutical composition of this invention.Accordingly, the terms “consisting essentially of” and “consisting of”are embodied in the term comprising.

As used herein, a “pharmaceutically acceptable” component is one that issuitable for use with humans and/or animals without undue adverse sideeffects (such as toxicity, irritation, and allergic response)commensurate with a reasonable benefit/risk ratio.

As used herein, the term “safe and effective amount” refers to thequantity of a component which is sufficient to yield a desiredtherapeutic response without undue adverse side effects (such astoxicity, irritation, or allergic response) commensurate with areasonable benefit/risk ratio when used in the manner of this inventionThe specific “safe and effective amount” will, obviously, vary with suchfactors as the particular condition being treated, the physicalcondition of the patient, the type of mammal being treated, the durationof the treatment, the nature of concurrent therapy (if any), and thespecific formulations employed and the structure of the compounds or itsderivatives.

As used herein, a “pharmaceutical addition salts” includes apharmaceutically acceptable salt of the anti-cancer compound. Theseinclude organic or inorganic acid salts of the amines. Preferred acidsalts are the hydrochlorides, acetates, salicylates, nitrates andphosphates.

As used herein, a “pharmaceutical carrier” is a pharmaceuticallyacceptable solvent, suspending agent or vehicle for delivering theanti-cancer agent to the animal or human. The carrier may be liquid orsolid and is selected with the planned manner of administration in mind.

As used herein, “cancer” refers to all types of cancers or neoplasm ortumors found in mammals, it includes leukemia.

As used herein, the “anti-cancer compounds” are N-chlorophenylcarbamatesand N-chlorophenylthiocarbamates.

As used herein, “viruses” includes viruses which cause diseases (viralinfection) in man and other warm blooded animals such as HIV virus,herpes, influenza and rhinoviruses.

As used herein “potentiators” are materials such as triprolidine and itscis-isomer which are used in combination with N-chlorophenylcarbamatesand N-chlorophenylthiocarbamates. Potentiators can affect the immunesystem or enhance the effectiveness of the drugs.

As used herein “chemotherapeutic agents” includes DNA-interactiveAgents, Antimetabolites, Tubulin-Interactive Agents, Hormonal agents andothers, such as Asparaginase or hydroxyurea.

B. The Anti-Cancer Compounds

The anti-cancer compounds are N-chlorophenylcarbamates andN-chlorophenylthiocarbamates which are known for their herbicidalactivities. They are systemic herbicides used to prevent and eradicatecertain plants or weeds. Systemic herbicides are differentiated fromother herbicides by their ability to be absorbed by the plant and tomove through the plant. This systemic ability is not a necessaryrequirement of the compounds of this invention.

The compounds have the following structure

wherein n is from 1 to 3, X is oxygen or sulfur and R is selected fromthe group consisting of hydrogen, lower alkyl and lower alkenyl,cyclohexyl, phenalkyl of up to 8 carbon atoms and phenyl, and thepharmaceutically acceptable salts of these compounds.

Preferred compounds are those in which R is alkyl with 1 to 4 carbons,preferably, isopropyl and X is oxygen, n is 1 and the chloro group is inthe 3 position on the phenyl group. N-3-chlorophenylcarbamate is a mostpreferred compound.

These compounds are prepared according to the method described in U.S.Pat. No. 2,695,225 issued to Witman (1954) and U.S. Pat. No. 2,734,911issued to Strain (1956).

C. Chemotherapeutic Agents

The chemotherapeutic agents are generally grouped as DNA-interactiveAgents, Antimetabolites, Tubulin-Interactive Agents, Hormonal agents andothers such as Asparaginase or hydroxyurea. Each of the groups ofchemotherapeutic agents can be further divided by type of activity orcompound. The chemotherapeutic agents used in combination withN-chlorophenylcarbamates and N-chlorophenylthiocarbamates includemembers of all of these groups. For a detailed discussion of thechemotherapeutic agents and their method of administration, see Dorr, etal, Cancer Chemotherapy Handbook, 2d edition, pages 15-34, Appleton &Lange (Connecticut, 1994) herein incorporated by reference.

DNA-interactive Agents include the alkylating agents, e.g. Cisplatin,Cyclophosphamide, Altretamine; the DNA strand-breakage agents, such asBleomycin; the intercalating topoisomerase II inhibitors, e.g.,Dactinomycin and Doxorubicin); the nonintercalating topoisomerase IIinhibitors such as, Etoposide and Teniposde; and the DNA minor groovebinder Plcamydin.

The alkylating agents form covalent chemical adducts with cellular DNA,RNA, and protein molecules and with smaller amino acids, glutathione andsimilar chemicals. Generally, these alkylating agents react with anucleophilic atom in a cellular constituent, such as an amino, carboxyl,phosphate, sulfhydryl group in nucleic acids, proteins, amino acids, orglutathione. The mechanism and the role of these alkylating agents incancer therapy is not well understood. Typical alkylating agentsinclude:

Nitrogen mustards, such as Chlorambucil, Cyclophosphamide, Isofamide,Mechlorethamine, Melphalan, Uracil mustard;

Aziridine such as Thiotepa

methanesulphonate esters such as Busulfan;

nitroso ureas, such as Carmustine, Lomustine, Streptozocin;

platinum complexes, such as Cisplatin, Carboplatin;

bioreductive alkylator, such as Mitomycin, and Procarbazine, Dacarbazineand Altretamine;

DNA strand breaking agents include Bleomycin;

DNA topoisomerase II inhibitors include the following:

Intercalators, such as Amsacrine, Dactinomycin, Daunorubicin,Doxorubicin, Idarubicin, and Mitoxantrone;

nonintercalators, such as Etoposide and Teniposide.

The DNA minor groove binder is Plicamycin.

The antimetabolites interfere with the production of nucleic acids byone or the other of two major mechanisms. Some of the drugs inhibitproduction of the deoxyribonucleoside triphosphates that are theimmediate precursors for DNA synthesis, thus inhibiting DNA replication.Some of the compounds are sufficiently like purines or pyrimidines to beable to substitute for them in the anabolic nucleotide pathways. Theseanalogs can then be substituted into the DNA and RNA instead of theirnormal counterparts. The antimetabolites useful herein include:

folate antagonists such as Methotrexate and trimeterxate

pyrimidine antagonists; such as Fluorouracil, Fluorodeoxyuridine, CB3717(N¹⁰-propargyl-5,8-dideazafolic acid), Azacitidine, Cytarabine, andFloxuridine;

purine antagonists include Mercaptopurine, 6-Thioguanine, Fludarabine,Pentostatin;

sugar modified analogs include Cyctrabine, Fludarabine;

ribonucleotide reductase inhibitors include hydroxyurea.

Tubulin Interactive agents act by binding to specific sites on tubulin,a protein that polymerizes to form cellular microtubules. Microtubulesare critical cell structure units. When the interactive agents bind onthe protein, the cell can not form microtubules Tubulin Interactiveagents include Vincristine and Vinblastine, both alkaloids andPaclitaxel.

Hormonal agents are also useful in the treatment of cancers and tumors.They are used in hormonally susceptible tumors and are usually derivedfrom natural sources. These include:

estrogens, conjugated estrogens and Ethinyl Estradiol andDiethylstilbesterol, Chlortrianisen and Idenestrol;

progestins such as Hydroxyprogesterone caproate, Medroxyprogesterone,and Megestrol;

androgens such as testosterone, testosterone propionate;fluoxymesterone, methyltestosterone;

Adrenal corticosteroids are derived from natural adrenal cortisol orhydrocortisone. They are used because of their anti inflammatorybenefits as well as the ability of some to inhibit mitotic divisions andto halt DNA synthesis. These compounds include, Prednisone,Dexamethasone, Methyiprednisolone, and Prednisolone.

Leutinizing hormone releasing hormone agents or gonadotropin-releasinghormone antagonists are used primarily the treatment of prostate cancer.These include leuprolide acetate and goserelin acetate. They prevent thebiosynthesis of steroids in the testes.

Antihormonal antigens include:

antiestrogenic agents such as Tamosifen,

antiandrogen agents such as Flutamide; and

antiadrenal agents such as AMitotane and Aminoglutethimide.

Hydroxyurea appears to act primarily through inhibition of the enzymeribonucleotide reductase.

Asparaginase is an enzyme which converts asparagine to nonfunctionalaspartic acid and thus blocks protein synthesis in the tumor.

Taxol is a preferred chemotherapeutic agent.

D. Potentiators

The “potentiators” can be any material which improves or increase theefficacy of the pharmaceutical composition or acts on the immune system.One such potentiator is triprolidine and its cis-isomer which are usedin combination with the chemotherapeutic agents and theN-chlorophenylcarbamates and N-chlorophenylthiocarbamates. Triprolidineis described in U.S. Pat. No. 5,114,951 (1992). Another potentiator isprocodazole, 1H-Benzimidazole-2-propanoic acid; [β-(2-benzimidazole)propionic acid; 2-(2-carboxyethyl)benzimidazole; propazol]. Procodazoleis a non-specific active immunoprotective agent against viral andbacterial infections and can be used with the compositions claimedherein. It is effective with the N-chlorophenylcarbamates and theN-chlorophenylthiocarbamates alone in treating cancers, tumors, leukemiaand viral infections or combined with chemotherapeutic agents.

Generally an amount effective to enhance the activity of thepharmaceutical composition is used.

Propionic acid and its salts and esters can also be used in combinationwith the pharmaceutical compositions claimed herein.

Antioxidant vitamins such as vitamins A, C and E and beta-carotene canbe added to these compositions.

E. Dosage

Any suitable dosage may be given in the method of the invention. Thetype of compound and the carrier and the amount will vary widelydepending on the species of the warm blooded animal or human, bodyweight, and tumor being treated. Generally a dosage of between about 2milligrams (mg) per kilogram (kg) of body weight and about 800 mg per kgof body weight is suitable. Preferably from 15 mg to about 500 mg/kg andmost preferably form about 15 mg/kg to about 150 mg/kg of body weight isused. Generally, the dosage in man is lower than for small warm bloodedmammals such as mice. A dosage unit may comprise a single compound ormixtures thereof with other compounds or other cancer inhibitingcompounds. The dosage unit can also comprise diluents, extenders,carriers and the like. The unit may be in solid or gel form such aspills, tablets, capsules and the like or in liquid form suitable fororal, rectal, topical or parenteral administration or intravenousadministration or by injection into or around the tumor site.

The range and ratio of N-chlorophenylcarbamates and theN-chlorophenylthiocarbamates to chemotherapeutic agent will depend onthe type of cancer or tumor being treated and the particularchemotherapeutic agent. The amount of chemotherapeutic agent used can belower than that of the N-chlorophenylcarbamates and theN-chlorophenylthiocarbamates and can range from 0.5 mg/kg body weight toabout 400 mg/kg body weight.

F. Dosage Delivery Forms

The chemotherapeutic agents, N-chlorophenylcarbamates and theN-chlorophenylthiocarbamates and, optionally, the potentiators aretypically mixed with a pharmaceutically acceptable carrier. This carriercan be a solid or liquid and the type is generally chosen based on thetype of administration being used. The active agent can becoadministered in the form of a tablet or capsule, liposome, as anagglomerated powder or in a liquid form. Examples of suitable solidcarriers include lactose, sucrose, gelatin and agar. Capsule or tabletscan be easily formulated and can be made easy to swallow or chew; othersolid forms include granules, and bulk .powders. Tablets may containsuitable binders, lubricants, diluents, disintegrating agents, coloringagents, flavoring agents, flow-inducing agents, and melting agents.Examples of suitable liquid dosage forms include solutions orsuspensions in water, pharmaceutically acceptable fats and oils,alcohols or other organic solvents, including esters, emulsions, syrupsor elixirs, suspensions, ,solutions and/or suspensions reconstitutedfrom non-effervescent granules and effervescent preparationsreconstituted from effervescent granules. Such liquid dosage forms maycontain, for example, suitable solvents, preservatives, emulsifyingagents, suspending agents, diluents, sweeteners, thickeners, and meltingagents. Oral dosage forms optionally contain flavorants and coloringagents. Parenteral and intravenous forms would also include minerals andother materials to make them compatible with the type of injection ordelivery system chosen.

Specific examples of pharmaceutical acceptable carriers and excipientsthat may be used to formulate oral dosage forms of the present inventionare described in U.S. Pat. No. 3,903,297 to Robert, issued Sep. 2, 1975.Techniques and compositions for making dosage forms useful in thepresent invention are described in the following references: 7 ModernPharmaceutics, Chapters 9 and 10 (Banker & Rhodes, Editors, 1979);Liebermnan et al., Pharmaceutical Dosage Forms: Tablets (1981); andAnsel, Introduction to Pharmaceutical Dosage Forms 2nd Edition (1976).

G. Method of Treatment

The method of treatment can be any suitable method which is effective inthe treatment of the particular virus or tumor type that is beingtreated. Treatment may be oral, rectal, topical, parenteral, intravenousadministration or injection into or around the tumor site and the like.The method of applying an effective amount also varies depending on thetumor being treated. It is believed that parenteral treatment byintravenous, subcutaneous, or intramuscular application. formulated withan appropriate carrier, additional cancer inhibiting compound orcompounds or diluent to facilitate application will be the preferredmethod of administering the compounds to warm blooded animals.

The method of treating viral infections may also be by oral, rectal,parenteral, topical or intravenous administration. The actual time anddosage will depend on the type of the virus being treated and thedesired blood levels.

The following examples are illustrative and are not meant to be limitingto the invention.

In addition to their combination with chemotherapeutic agents andpotentiators, N-chlorophenylcarbamates and theN-chlorophenylthiocarbamates can be combined with fungicides, herbicidesor other antiviral agents. Preferred herbicides and fungicides includecarbendazim, fluoconazole, benomyl, glyphosate and propicodazole.

The N-chlorophenylcarbamates and N-chlorophenylthiocarbamates are alsoeffective against viruses including rhinovirus, HIV, herpes, andinfluenza.

In the treatment of viral infections, the N-chlorophenylcarbamates andthe N-chlorophenylthiocarbamates can be combined with other anti-viralagents to effectively treat viral infections.

HIV Chronic Study

In this model, Chloropropham showed 79% suppression of HIV replicationin monocytes. The positive control, interferon, showed 80% suppression.There was 101% increase in HIV replication in T-cells compared to 60%suppression for interferon. AZT showed no action in this model.

Chronic HIV-1 infected cells U1 were derived from an acute HIV-1infection of the promonocytic cell line, U937. The chronic HIV-1infected cells, ACH-2 were derived from an acute HIV-1 infection of theT cell line, A3.01.

These cells were cultured in medium and the phorbol ester, PMA. PMAcauses the cells (both U1 and ACH-2) to be activated and not divide butit also causes the U-1 cells to differentiate. This results in fewercells in the PMA-treated cultures than the media alone cultures. Cellviability was measured when these cell lines were treated with the testcompounds.

Both cell lines constituitively produce a small amount of HIV-1. ACH-2cell lines tend to produce more HIV-1 than U1 cells as shown by p-24ELISA. When either cell line is cultured in the presence of PMA there isan increase in the quantity of HIV-1 produced as measured by the p-24antigen ELISA.

In addition, the number of institute positive HIV mRNA expressing cellsper microscopic field is measured. Comparisons can be made from thesenumbers since the same number of cells were adhered to the glass slidesfor each drug concentration (10×10⁶ cells/ml).

What is claimed is:
 1. A pharmaceutical composition for treating cancersor tumors susceptible to treatment comprising: a safe and effectiveamount of a chemotherapeutic agent wherein said chemotherapeutic agentis selected from the group consisting of Methotrexate, Fluorouracil,Fluorodeoxyuridine, N¹⁰-propargyl-5,8-dideazafolic acid, Azacitidine,Cytarabine, Floxuridine, Mercaptopurine, 6-Thioguanine, Pentostatin,Fludarabine, Tamoxifen and Busulfan; and a safe and enhanced amount ofan N-chlorophenylcarbamate or N-chlorophenylthiocarbamate of theformula:

wherein n is from 1 to 3; X is selected from the group consisting oxygenand sulfur; and R is selected from the group consisting of hydrogen,lower alkyl, lower alkenyl, cyclohexyl, phenyl, and phenalkyl of up to 8carbon atoms; or a pharmaceutically acceptable inorganic or organic acidsalt thereof.
 2. A pharmaceutical composition according to claim 1comprising a pharmaceutically acceptable carrier and a safe and enhancedamount of N-chlorophenylcarbamates and N-chloraphenylthiocarbamates. 3.A pharmaceutical composition according to claim 2 wherein saidpharmaceutically acceptable salt is selected from the group consistingof hydrochlorides, acetates, salicylates, nitrates, phosphate salts andmixtures thereof.
 4. The pharmaceutical composition of claim 1 whereinthe chemotherapeutic agent is Busulfan, and wherein theN-chlorophenylcarbamate is N-3-chlorophenylcarbamate.
 5. Thepharmaceutical composition of claim 1 wherein the chemotherapeutic agentis Methotrexate, and wherein the N-chlorophenylcarbamate isN-3-chlorophenylcarbamate.
 6. The pharmaceutical composition of claim 1wherein the chemotherapeutic agent is Fluorouracil, and wherein theN-chlorophenylcarbamate is N-3-chlorophenylcarbamate.
 7. Thepharmaceutical composition of claim 1 wherein the chemotherapeutic agentis azacitidine, and wherein the N-chlorophenylcarbamate isN-3-chlorophenylcarbamate.
 8. The pharmaceutical composition of claim 1wherein the chemotherapeutic agent is cytarabine, and wherein theN-chlorophenylcarbamate is N-3-chlorophenylcarbamate.
 9. Thepharmaceutical composition of claim 1 wherein the chemotherapeutic agentis pentostatin, and wherein the N-chlorophenylcarbamate isN-3-chlorophenylcarbamate.
 10. The pharmaceutical composition of claim 1wherein the chemotherapeutic agent is tamoxifen, and wherein theN-chlorophenylcarbamate is N-3-chlorophenylcarbamate.
 11. A method oftreating cancer susceptible to treatment comprising administering a safeand effective amount of a pharmaceutical composition to a patient inneed thereof comprising; a safe and effective amount of achemotherapeutic agent wherein said chemotherapeutic agent is selectedfrom the group consisting of Methotrexate, Fluorouracil,Fluorodeoxyuridine, N¹⁰-propargyl-5,8-dideazafolic acid, Azacitidine,Cytarabine, Floxuridine, Mercaptopurine, 6-Thioguanine, Pentostatin,Fludarabine, Tamoxifen and Busulfan; and a safe and enhanced amount ofan N-chlorophenylcarbamate or N-chlorophenylthiocarbamate of theformula:

wherein n is from 1 to 3, X is selected from the group consisting ofoxygen and sulfur, and R is selected from the group consisting ofhydrogen, lower alkyl, lower alkenyl, cyclohexyl, phenalkyl of up to 8carbon atom, and phenyl, or a pharmaceutically acceptable organic orinorganic acid salt thereof.
 12. The method of claim 11 wherein thechemotherapeutic agent is Busulfan, and wherein theN-chlorophenylcarbamate is N-3-chlorophenylcarbamate.
 13. A method oftreating cancer according to claim 11 comprising administering a safeand effective amount of an N-chlorophenylcarbamate wherein R is alkyl offrom 1 to 4 carbons, n is 1, X is oxygen and the chloro is in the 3position of the phenyl.
 14. A method according to claim 13 wherein from2 mg/kg body weight to 400 mg/kg of said N-chlorophenylcarbamate isadministered.
 15. A method according to claim 14 wherein saidpharmaceutical composition is administered orally or enterically,intravenously, parenterally or by injection into or around the cancer.16. A method according to claim 15 wherein from 0.5 mg/kg body weight to400 mg/kg body weight of said chemotherapeutic agent is administered.17. A composition effective for treating cancers or tumors susceptibleto treatment comprising: a safe and effective amount of achemotherapeutic agent wherein said chemotherapeutic agent is aDNA-interactive Agent; and a safe and enhanced amount of anN-chlorophenylcarbamate or N-chlorophenylthiocarbamate of the formula:

wherein X is selected from the group consisting of oxygen and sulfur, nis from 1 to 3, and R is selected from the group consisting of hydrogen,lower alkyl, lower alkenyl, cyclohexyl, phenalkyl of up to 8 carbonatoms, and phenyl, or a pharmaceutically acceptable organic or inorganicacid salt thereof, and a safe and effective carrier.